Wet preservation of tissue

ABSTRACT

A system and method are provided for wet storage of tissue. In an embodiment, a solution for the wet preservation of tissue may include between about 0.1% to about 50% by volume dimethyl sulfoxide (DMSO) and one or more soluble monovalent or divalent metal cationic salts. A wet-preserved tissue and method for preparing the wet-preserved tissue for ultimate use, is also provided.

TECHNICAL FIELD

The present disclosure generally relates to the preservation of tissue,and more particularly relates to the preservation of tissue using aliquid medium.

BACKGROUND

Biological tissues may often provide improved functional performance ascompared to equivalent synthetic devices when used in in vivoimplantation. However, the availability and usage of tissue grafts maybe restricted by inherent supply constraints and logistic concerns ofharvest, transportation, and storage. Various techniques have beendeveloped that allow for the harvesting, delivery, and storage oftissues that may be suitable for use in clinical implantation. However,many of the developed techniques may require the use of specializedrefrigeration equipment for storing and shipping the donor tissue atextremely low temperatures, e.g., less than −40 degrees C., and, moreparticularly, −80 degrees C. Such requirements may frequently restrictthe facilities that are able to make use of the available tissues, andmay complicate the transportation of tissue to end-use clinicalfacilities. It is also often more desirable to be able to work withwet-preserved tissue, than dry-preserved tissue which requires addedsteps and pre-operation prep time, including for rehydration of thedried tissue. Some implementations consistent with the presentdisclosure may overcome and/or reduce such problems to provide a morereadily usable wet-preserved tissue.

SUMMARY

According to an implementation, a solution for the wet preservation oftissue may include between about 0.1% to about 50% by volume dimethylsulfoxide (DMSO). The solution may also include one or more solublesalts of sodium, potassium, calcium, magnesium or other monovalent ordivalent metal cationic salts.

One or more of the following features may be included. The solution mayinclude between about 2% to about 15% by volume DMSO. The solution mayinclude greater than about 2% by volume DMSO. The solution may includegreater than about 5% by volume DMSO. The solution may include greaterthan about 10% by volume DMSO.

The one or more soluble salts may provide one or more of between about43 mM and about 2.6 M sodium cation, about 2.57 M sodium cation, betweenabout 2.7 mM and about 5.4 mM potassium cation, between about 0.9 mM andabout 2.7 mM calcium cation, and/or between about 0.945 mM and about 1.2mM magnesium cation. The one or more soluble salts of sodium may includebetween about 2.5 g/L to about 150 g/L sodium chloride. The one or moresoluble salts of potassium may include between about 0.2 g/L to about0.4 g/L potassium chloride. The one or more soluble salts of calcium mayinclude between about 0.1 g/L to about 0.3 g/L calcium chloride. The oneor more soluble salts of magnesium may include between about 0.09 toabout 0.11 g/L magnesium chloride. The one or more soluble salts ofsodium may include between about 0.2 to about 0.8 g/L sodiumbicarbonate.

The solution may include between about 2% to about 15% DMSO by volume.The one or more soluble salts provide one or more of between about 43 mMand about 2.6 M sodium cation, between about 2.7 mM and about 5.4 mMpotassium cation, between about 0.9 mM and about 2.7 mM calcium cation,and/or between about 0.945 mM and about 1.2 mM magnesium cation.

According to another implementation, a method of preserving tissue mayinclude preparing a specimen of tissue. The specimen of tissue may bestored in a solution including between about 0.1% to about 50% by volumeDMSO. The solution may also include one or more soluble salts of:sodium, potassium, calcium, magnesium, or other monovalent or divalentmetal cationic salts, particularly of alkali metal and alkaline earthmetal cations.

One or more of the following features may be included. The specimen oftissue may be nerve tissue. The nerve tissue may be one of humanallograft nerve tissue and/or animal nerve tissue. The specimen oftissue may include one or more of human vascular tissue, urologicaltissue, tendons, and muscle tissue. The specimen of tissue may includeone or more of animal vascular tissue, urological tissue, tendons, andmuscle tissue. Preparing the specimen of tissue may includedecellularizing the tissue.

The solution may include between about 2% to about 15% by volume DMSO.The solution may include greater than about 2% by volume DMSO. Thesolution may include greater than about 5% by volume DMSO. The solutionmay include greater than about 10% by volume DMSO.

The one or more soluble salts may provide one or more of between about43 mM and about 2.6 M sodium cation, between about 2.7 mM and about 5.4mM potassium cation, between about 0.9 mM and about 2.7 mM calciumcation, and/or between about 0.945 mM and about 1.2 mM magnesium cation.The one or more soluble salts of sodium may include between about 2.5g/L and about 150 g/L sodium chloride. The one or more soluble salts ofpotassium may include between about 0.2 g/L to about 0.4 g/L potassiumchloride. The one or more soluble salts of calcium may include betweenabout 0.1 g/L to about 0.3 g/L calcium chloride. The one or more solublesalts of magnesium may include between about 0.09 to about 0.11 g/Lmagnesium chloride. The one or more soluble salts of sodium may includebetween about 0.2 g/L to about 0.8 g/L sodium bicarbonate.

The solution may include between about 2% to about 15% DMSO by volume.The one or more soluble salts may provide one or more of between about43 mM and about 2.6 M sodium cation, between about 2.7 mM and about 5.4mM potassium cation, between about 0.9 mM and about 2.7 mM calciumcation, and/or between about 0.945 mM and about 1.2 mM magnesium cation.

According to another implementation, a wet-preserved tissue may includea tissue specimen disposed in a solution including between about 0.1% toabout 50% by volume DMSO. The solution may also include one or moresoluble salts. The one or more soluble salts may provide one or more ofbetween about 43 mM and about 2.6 M sodium cation, between about 2.7 mMand about 5.4 mM potassium cation, between about 0.9 mM and about 2.7 mMcalcium cation, and/or between about 0.945 mM and about 1.2 mM magnesiumcation.

One or more of the following features may be included. The one or moresoluble salts may include one or more of between about 2.5 g/L to about150 g/L sodium chloride, between about 0.2 g/L to about 0.4 g/Lpotassium chloride, between about 0.1 g/L to about 0.3 g/L calciumchloride, between about 0.2 g/L to about 0.8 g/L sodium bicarbonate,and/or between about 0.09 g/L to about 0.11 g/L magnesium chloride.

The specimen of tissue may be nerve tissue. The nerve tissue may be oneof human nerve tissue and/or animal nerve tissue. The specimen of tissuemay include one or more of human vascular tissue, urological tissue,tendons, and/or muscle tissue. The specimen of tissue may include one ormore of animal vascular tissue, urological tissue, tendons, and/ormuscle tissue. Preparing the specimen of tissue may includedecellularizing the tissue. The wet-preserved tissue specimen mayultimately be implanted in a human or in an animal.

The solution may include between about 2% to about 15% by volume DMSO.The solution may include greater than about 2% by volume DMSO. Thesolution may include greater than about 5% by volume DMSO. The solutionmay include greater than about 10% by volume DMSO. The tissue may be atleast partially immersed in the solution.

According to another implementation, a method of using a wet-preservedtissue specimen may include removing the specimen of tissue which hadbeen disposed in a solution including between about 0.1% to about 50% byvolume DMSO. The solution may also include one or more soluble salts.The one or more soluble salts may provide one or more of between about43 mM and about 2.6 M sodium cation, between about 2.7 mM and about 5.4mM potassium cation, between about 0.9 mM and about 2.7 mM calciumcation, and/or between about 0.945 mM and about 1.2 mM magnesium cation.The method may also include reducing the amount of DMSO within thetissue to a residual amount below a predetermined threshold.

One or more of the following features may be included. The one or moresoluble salts may include one or more of between about 2.5 g/L to about150 g/L sodium chloride, between about 0.2 g/L to about 0.4 g/Lpotassium chloride, between about 0.1 g/L to about 0.3 g/L calciumchloride, between about 0.2 g/L to about 0.8 g/L sodium bicarbonate,and/or between about 0.09 g/L to about 0.11 g/L magnesium chloride.

The specimen of tissue may be nerve tissue. The nerve tissue may be oneof human nerve tissue and/or animal nerve tissue. The specimen of tissuemay include one or more of human vascular tissue, urological tissue,tendons, and/or muscle tissue. The specimen of tissue may include one ormore of animal vascular tissue, urological tissue, tendons, and/ormuscle tissue. Preparing the specimen of tissue may includedecellularizing the tissue.

The solution may include between about 2% to about 15% by volume DMSO.The solution may include greater than about 2% by volume DMSO. Thesolution may include greater than about 5% by volume DMSO. The solutionmay include greater than about 10% by volume DMSO.

While the amount of DMSO within the tissue, as packaged, may exceed 50mg, the residual amount of DMSO within the tissue may be at or belowabout 50 mg prior to implantation.

DETAILED DESCRIPTION OF EXAMPLE EMBODIMENTS

In this disclosure, the term “based on” means “based at least in parton.” The singular forms “a,” “an,” and “the” include plural referentsunless the context dictates otherwise. The term “exemplary” is used inthe sense of “example” rather than “ideal.” The terms “comprises,”“comprising,” “includes,” “including,” or other variations thereof, areintended to cover a non-exclusive inclusion such that a process, method,or product that comprises a list of elements does not necessarilyinclude only those elements, but may include other elements notexpressly listed or inherent to such a process, method, article, orapparatus. The relative term “about,” is used to indicate a possiblevariation of ±10% of a stated or understood value. In addition, the term“between” used in describing ranges of values is intended to include theminimum and maximum values described herein.

In general, embodiments consistent with the present disclosure mayrelate to systems and methods for preserving tissue. For example, thedisclosed systems and methods may allow tissue specimens to be preservedwhile maintaining structural integrity and/or functional stability ofthe tissue. In some implementations, maintaining the structuralintegrity and/or functional stability of the tissue may allow successfulin vivo implantation of the tissue. Consistent with example embodiments,a tissue specimen to be preserved may be stored in a solution includingless than about 50% DMSO and a selection of mono and/or divalent cationsand/or salts. In some implementations, storage of tissue specimens insolutions described herein may allow for prolonged room temperaturestorage, and/or storage at common refrigeration temperatures, of tissuewhile maintaining structural integrity and/or functional stability ofthe tissue.

Consistent with the present disclosure a variety of tissue types may bepreserved. For example, according to an illustrative example embodiment,the disclosed systems and methods may be utilized for preserving nervetissue, e.g., which may be subsequently used for in vivo implantation(and/or any other suitable use). In various additional and/oralternative embodiments, the disclosed systems and methods may beutilized for preserving other tissues, including, but not limited to,vascular tissue, urological tissue (e.g., bladder wall, ureter tissue,urethra tissue), skeletal tissue (e.g., bone, cartilage, ligaments,fascia, tendons), muscle tissue (e.g., striated and/or smooth muscletissue), skin, dura, myocardium, neural tissue, intestine, gingiva, andthe like. In some example embodiments, the tissue may include harvestedhuman nerve allograft tissue, e.g., which may be intended for use inclinical repair and/or restoration procedures. It will be appreciatedthat, in addition and/or as an alternative to human nerve allografttissue, the systems and methods herein may also be utilized inconnection with animal nerve tissue, as well as various other humanand/or animal tissues. According to an illustrative example, animaltissue, whether nerve tissue or another type of tissue, may includetissue from a ruminant.

In some implementations, a tissue specimen may be prepared forpreservation through a variety of processes that may isolate one or morespecific tissue types based upon, for example, an intended end use ofthe tissue. For example, in an illustrative example embodiment in whichthe tissue may include nerve tissue intended for in vivo implantation.In such an embodiment, the tissue may be harvested from a donor (such asa cadaver), and may be cleaned and processed to isolate the desiredtissue. In an example embodiment, harvested human nerve tissue may becleaned (e.g., to remove fat and/or other extraneous, non-nerve tissueand components, including antigens). Further, the harvested nerve tissuemay undergo various chemical and enzymatic processes to decellularizethe tissue. It will be appreciated that a variety of processes may beutilized to clean and decellularize the tissue specimens, and, as such,the present disclosure is not intended to be limited to any specificpreparation process. One suitable illustrative methodology for preparinga tissue specimen for preservation is described in U.S. Pat. No.9,572,911, entitled “Method for Decellularization of Tissue Grafts,” theentire disclosure of which is incorporated herein by reference. It willbe appreciated, however, that various other methods for preparing tissuespecimens may be used.

Once a tissue specimen has been prepared for storage in a desired manner(e.g., via cleaning and/or decellularization and/or any other processingsteps), the tissue specimen may be disposed in a preservation solutionthat may permit extended storage of the tissue, while maintaining thestructural integrity and/or functional stability of the tissue. Ingeneral, the present disclosure may provide a wet storage approach,whereby the tissue may be disposed in the preservation solution for theduration of the tissue storage. In some implementations, the tissue maybe disposed in the preservation solution for the duration of the tissuestorage by packaging the tissue in a reservoir (e.g., which may includeany suitable liquid containment vessel) that may be at least partiallyfilled with the preservation solution. Consistent with such animplementation, the tissue disposed in the preservation solution mayremain at least partially immersed in the preservation solution for theduration of the tissue storage. In some implementations, the tissue maybe disposed in the preservation solution by saturating the tissue withthe preservation solution. The tissue saturated with the preservationsolution may then be sealed in a suitable packaging arrangement that mayprevent and/or reduce the rate of drying of the tissue during storage.In such an implementation, the saturated tissue may be disposed in thepreservation solution, but the packaging may include relatively little,or no, free preservation solution outside of saturated tissue itself(e.g., as compared to an embodiment in which the tissue may be at leastpartially immersed in preservation solution that may at least partiallyfill the reservoir and/or other liquid containment vessel). The tissuemay be saturated with the preservation solution in any suitable manner,such as by being immersed in the preservation solution for an adequateperiod of time to achieve a desired level of saturation.

As generally discussed above, consistent with some embodiments, thetissue may be stored in a liquid containment vessel that may retain thetissue in an enclosed volume of the preservation solution. In someembodiments, the volume of preservation solution surrounding the tissuemay be minimized. For example, the tissue may be saturated with thepreservation solution and subsequently contained in a liquid containmentvessel that may provide limited, or no, excess liquid volume surroundingthe tissue specimen. It will be appreciated that various implementationsutilizing different liquid containment vessels and different retainedvolumes of preservation solution (e.g., in relation to the size and/orvolume of the tissue specimen) may be achieved. Examples of suitableliquid containment vessels may include, but are not limited to, sealedvials, sealed foil pouches, plastic blister packaging with peelableand/or rupturable film or foil seal, or any other suitable liquidcontainment vessel.

In some implementations, the tissue specimen may be sterilized, e.g.,either before or after being placed in the preservation solution. In anexample embodiment, the tissue specimen may be sealed in a liquidcontainment vessel, including the preservation solution, and maysubsequently be sterilized using gamma radiation, e.g., in a generallyknown manner. In some implementations, sterilization via gamma radiationmay be carried out at low temperature, e.g., a temperature on the orderof −80 degrees C., although such parameters may vary depending upon theexact sterilization process utilized. Further in some embodiments, priorto in vivo implantation, an amount of one or more components of thepreservation solution in the tissue specimen, may be reduced to or belowa predetermined threshold to provide the desired residual amount of thatcomponent in the tissue specimen. For example, in some embodiments thetissue specimen may undergo one or more soaking and/or rinsingprocesses, with or without agitation, e.g., to decrease the amount ofany residual preservation solution to, or below, a desired threshold.Consistent with such an example, at the time of in vivo implantation,the tissue specimen may be removed from the liquid containment vessel,and may be placed in a rinse basin including a suitable rinse solution,such as Lactated Ringers Solution (“LRS”), saline, phosphate bufferedsaline (“PBS”), or other suitable rinse solution/liquid. In someimplementations more than one rinse solution may be utilized and/or morethan one soak and/or rinse operation may be performed prior toimplantation of the tissue specimen. In some implementations, one ormore soak and/or rinse operations may be carried out to decrease theDMSO amount in the tissue specimen to be at or below a desired thresholdresidual amount. In an illustrative example embodiment, the tissuespecimen may be rinsed to reduce the DMSO amount in the tissue specimento be at or below 50 mg prior to implantation. For example, in anillustrative example rinse protocol, the tissue specimen may be washedfor ten minutes with 100 mL of a 0.9% sodium chloride rinse solution(e.g., a normal saline solution) to reduce the DMSO amount to be at orbelow 50 mg. It will be appreciated that other soak and/or rinseprotocols may be utilized to provide a tissue specimen having adifferent residual amount of DMSO. In some implementations, one or morerinse operations may be carried out to decrease the amount of one ormore other components of the preservation solution in the tissuespecimen to or below a desired residual threshold amount.

As discussed above, consistent with various embodiments, wet storage oftissue specimens in a preservation solution consistent with the presentdisclosure may allow prolonged storage of tissue while maintainingstructural integrity and/or functional stability of the tissue. Forexample, decellularized human nerve tissue stored in a preservationsolution consistent with the present disclosure has exhibited favorablestability as evaluated using neurite outgrowth bioassay and histologicalassessment of the endoneurial tube assessment) based on the structure ofindividual endoneurial tubes within the tissue specimen.

As noted, in some example implementations, structural integrity andfunctional stability of tissue specimens may be maintained overprolonged storage periods within preservation solutions consistent withthe present disclosure. For example, in some implementations,preservation solutions consistent with the present disclosure may becapable of achieving a useful shelf life for stored tissues on the orderof about 2 years. It will be appreciated that, depending upon tissuetype, storage conditions, and exact solution formulations, longer orshorter useful shelf lives may be achieved. In some implementations, thestructural integrity and/or functional stability of tissue specimens maybe achieved using preservation solutions herein without the use ofextreme low temperature storage. For example, some conventional tissuestorage approaches may utilize low temperatures as part of the storageprocess. For example, some storage processes may utilize storagetemperatures on the order of about −80 degrees C. In some embodiments, apreservation solution consistent with the present disclosure may allowprolonged storage of tissue at room temperature, and/or at refrigerationlevels achievable with readily available conventional refrigerationsystems. Consistent with various embodiments, prolonged storage oftissue may be achieved at temperatures in the range of about 10 degreesC. to about 30 degrees C., and, more particularly, 15 degrees C. toabout 30 degrees C., while maintaining structural integrity and/orfunctional stability of the tissue. According to other implementations,lower or higher storage temperatures may be utilized. The ability toachieve prolonged storage of tissue at such temperatures may be usefulfor clinical environments that may not have access to specializedrefrigeration equipment capable of extremely low storage temperatures.Similarly, the ability to achieve prolonged storage of tissue at suchtemperatures may facilitate shipment of tissue (e.g., from a productionfacility that may prepare the tissue specimens to an end-useenvironment, such as a clinical facility), without the requirement forspecialized cold transport.

In an example embodiment, a solution for the wet preservation of tissuemay include a DMSO solution in combination with various salts that may,for example, allow for prolonged storage of a tissue specimen whilemaintaining structural integrity and/or functional stability of thetissue. In an embodiment, the solution may include less than about 50%DMSO by volume of the solution. In some example embodiments the solutionmay include between about 0.1% to about 50% DMSO by volume. In someexample embodiments, the solution may include less than about 15% DMSOby volume. In some implementations, the solution may include greaterthan about 2% DMSO by volume of the solution. According to otherillustrative examples, the solution may include any concentration ofDMSO between about 2% to about 15% DMSO by volume of the solution. Forexample, the solution may include about 5% DMSO by volume of thesolution, 10% by volume of the solution, as well as any otherincremental value between about 2% to about 15% DMSO by volume of thesolution. Further, in various implementations, the solution may include,for example, between about 2% to about 10% DMSO by volume of thesolution. In some implementations, the solution may include betweenabout 2% to about 5% DMSO by volume of the solution. In someimplementations, the solution may include between about 5% to about 15%DMSO by volume of the solution. In some implementations, the solutionmay include between about 10% to about 15% DMSO by volume of thesolution. In some implementations, the solution may include betweenabout 5% to about 10% DMSO by volume of the solution.

As generally discussed above, the wet preservation solution may includeone or more of various monovalent and/or divalent cations and/or salts(e.g., that may provide various monovalent and/or divalent cations insolution). As it is recognized that each of the salts appropriate forthis disclosure will dissociate, as used herein, the term “solution” or“wet preservation solution” is therefore used to refer to and include asolution of DMSO and the soluble salts used to form the solution, aswell as a solution of DMSO and the independent monovalent and/ordivalent cations and/or anions provided from the dissociation of thosesalts. For example, the solution may include one or more differentmonovalent and/or divalent metal cations, such as, but not limited to,sodium, potassium, calcium, magnesium, and/or zinc. In one embodiment,the solution may include one or more of sodium chloride, potassiumchloride, calcium chloride, sodium bicarbonate, and/or magnesiumchloride as contributors of the one or more cations. It will beappreciated that in other embodiments, different salts may be utilizedas cation contributors. In an illustrative example, the one or moresoluble salts may provide one or more of between about 43 mM and about2.6 M sodium cation, between about 2.7 mM and about 5.4 mM potassiumcation, between about 0.9 mM and about 2.7 mM calcium cation, and/orbetween about 0.945 mM and about 1.2 mM magnesium cation.

In some illustrative example embodiments, the solution may includebetween about 2.5 g/L and about 150 g/L sodium chloride. In some exampleembodiments, the solution may include between about 2.5 g/L and about 12g/L sodium chloride. In some example embodiments, the solution mayinclude between about 2.5 g/L and about 6 g/L sodium chloride. In someexample embodiments, the solution may include between about 6 g/L andabout 12 g/L sodium chloride.

In some illustrative example embodiments, the solution may includebetween about 0.2 g/L to about 0.4 g/L potassium chloride. In anillustrative example embodiment, the solution may include about 0.3 g/Lpotassium chloride. In some illustrative example embodiments, thesolution may include between about 0.1 g/L to about 0.3 g/L calciumchloride. In an illustrative example embodiment, the solution mayinclude about 0.2 g/L calcium chloride. In some illustrative exampleembodiments, the solution may include between about 0.09 g/L to about0.11 g/L magnesium chloride. In an illustrative example embodiment, thesolution may include about 0.1 g/L magnesium chloride. In someillustrative example embodiments, the solution may include between about0.2 g/L to about 0.8 g/L sodium bicarbonate. In an illustrative example,the solution may include about 0.4 g/L sodium bicarbonate. In variousembodiments, one or more of the foregoing salts and/or cations donatedby the foregoing salts, may be omitted from the solution. Further, insome embodiments, one or more additional salts and/or cations may beincluded.

While some of the foregoing example embodiments have been described inthe context of preserving nerve tissue for use in nerve graftapplications, it will be appreciated that the principles, features,and/or advantages described herein may be equally applicable to othertypes of tissues and/or tissues for other intended applications. It willalso be appreciated that the solutions, tissues, and preservationmethodologies may be susceptible to various modifications, variations,and substitutions. Accordingly, the present disclosure should not belimited by any of the disclosed example embodiments, and should beafforded the full scope of the appended claims.

What is claimed is:
 1. A composition including a liquid and a solid,wherein the composition comprises: a wet-preserved nerve configured forimplantation into a subject; and a liquid wet-preservation solution,comprising: between about 0.1% to about 50% by volume dimethyl sulfoxide(DMSO); and one or more soluble salts, wherein the one or more solublesalts include one or more of: between about 2.5 q/L and about 150 g/Lsodium chloride; between about 0.2 q/L to about 0.4 g/L potassiumchloride; between about 0.1 q/L to about 0.3 g/L calcium chloride;between about 0.2 q/L to about 0.8 g/L sodium bicarbonate; or betweenabout 0.09 q/L to about 0.11 q/L magnesium chloride.
 2. The compositionaccording to claim 1, wherein the one or more soluble salts provide oneor more of: between about 43 mM and about 2.6 M sodium cation, betweenabout 2.7 mM and about 5.4 mM potassium cation, between about 0.9 mM andabout 2.7 mM calcium cation, or between about 0.945 mM and about 1.2 mMmagnesium cation.
 3. The composition according to claim 1, wherein thenerve is one of a human nerve or an animal nerve.
 4. The compositionaccording to claim 1, wherein the nerve is decellularized.
 5. Thecomposition according to claim 1, wherein the liquid wet-preservationsolution includes between about 2% to about 15% by volume DMSO.
 6. Thecomposition according to claim 1, wherein the liquid wet-preservationsolution includes greater than about 2% by volume DMSO to about 50% byvolume DMSO.
 7. The composition according to claim 1, wherein the liquidwet-preservation solution includes greater than about 5% by volume DMSOto about 50% by volume DMSO.
 8. The composition according to claim 1,wherein the liquid wet-preservation solution includes greater than about10% by volume DMSO to about 50% by volume DMSO.
 9. The compositionaccording to claim 1, wherein the wet-preserved nerve is saturated inthe liquid wet-preservation solution.
 10. The composition according toclaim 1, wherein the one or more soluble salts include one or more of:between about 2.5 g/L and about 12 g/L sodium chloride; about 0.3 g/Lpotassium chloride; about 0.2 g/L calcium chloride; about 0.1 g/Lmagnesium chloride; or about 0.4 g/L sodium bicarbonate.
 11. Thecomposition according to claim 1, wherein the one or more soluble saltsinclude multiple soluble salts, and wherein the multiple soluble saltsinclude: between about 2.5 g/L and about 12 g/L sodium chloride; about0.3 g/L potassium chloride; about 0.2 g/L calcium chloride; about 0.1g/L magnesium chloride; and about 0.4 g/L sodium bicarbonate.
 12. Acomposition including a liquid and a solid, wherein the compositioncomprises: a wet-preserved nerve configured for implantation into asubject; and a liquid wet-preservation solution consisting of: betweenabout 0.1% to about 50% by volume dimethyl sulfoxide (DMSO); and one ormore soluble salts, wherein the one or more soluble salts include one ormore of: between about 2.5 q/L and about 150 g/L sodium chloride;between about 0.2 q/L to about 0.4 g/L potassium chloride; between about0.1 q/L to about 0.3 g/L calcium chloride; between about 0.2 q/L toabout 0.8 g/L sodium bicarbonate; or between about 0.09 q/L to about0.11 q/L magnesium chloride; wherein the composition has a shelf life ofup to about 2 years.
 13. The composition according to claim 12, whereinthe one or more soluble salts provide one or more of: between about 43mM and about 2.6 M sodium cation, between about 2.7 mM and about 5.4 mMpotassium cation, between about 0.9 mM and about 2.7 mM calcium cation,or between about 0.945 mM and about 1.2 mM magnesium cation.
 14. Thecomposition according to claim 1, wherein the composition has a shelflife of up to about 2 years.
 15. A method comprising: removing awet-preserved nerve configured for implantation into a subject from aliquid wet-preservation solution comprising: between about 0.1% to about50% by volume dimethyl sulfoxide (DMSO); and one or more soluble salts,wherein the one or more soluble salts include one or more of: betweenabout 2.5 q/L and about 150 g/L sodium chloride; between about 0.2 q/Lto about 0.4 g/L potassium chloride; between about 0.1 q/L to about 0.3g/L calcium chloride; between about 0.2 q/L to about 0.8 g/L sodiumbicarbonate; or between about 0.09 q/L to about 0.11 q/L magnesiumchloride; and reducing the amount of DMSO within the wet-preserved nerveto an amount at or below a predetermined threshold.
 16. The methodaccording to claim 15, wherein the one or more soluble salts provide oneor more of: between about 43 mM and about 2.6 M sodium cation, betweenabout 2.7 mM and about 5.4 mM potassium cation, between about 0.9 mM andabout 2.7 mM calcium cation, or between about 0.945 mM and about 1.2 mMmagnesium cation.
 17. The method according to claim 15, wherein thewet-preserved nerve is one of a human nerve or an animal nerve.
 18. Themethod according to claim 15, further comprising decellularizing thenerve prior to disposing the nerve in the liquid wet-preservationsolution.
 19. The method according to claim 15, wherein the liquidwet-preservation solution includes between about 2% to about 15% byvolume DMSO.
 20. The method according to claim 15, wherein the liquidwet-preservation solution includes greater than about 2% by volume DMSOto about 50% by volume DMSO.
 21. The method according to claim 15,wherein the liquid wet-preservation solution includes greater than about5% by volume DMSO to about 50% by volume DMSO.
 22. The method accordingto claim 15, wherein the liquid wet-preservation solution includesgreater than about 10% by volume DMSO to about 50% by volume DMSO. 23.The method according to claim 15, wherein the predetermined threshold is50 mg, and the amount of DMSO within the wet-preserved nerve is reducedto at or below 50 mg DMSO.
 24. The method according to claim 15, whereinthe one or more soluble salts include one or more of: between about 2.5g/L and about 12 g/L sodium chloride; about 0.3 g/L potassium chloride;about 0.2 g/L calcium chloride; about 0.1 g/L magnesium chloride; orabout 0.4 g/L sodium bicarbonate.
 25. The method according to claim 15,wherein the one or more soluble salts include multiple soluble salts,and wherein the multiple soluble salts include: between about 2.5 g/Land about 12 g/L sodium chloride; about 0.3 g/L potassium chloride;about 0.2 g/L calcium chloride; about 0.1 g/L magnesium chloride; andabout 0.4 g/L sodium bicarbonate.
 26. The method of claim 15, furthercomprising: sterilizing the nerve at a temperature of −80 degrees C.;and increasing the temperature of the nerve to above freezing oncesterilization is complete.